This project will apply techniques for fast time-resolved magnetic circular dichroism (TRMCD) and magnetic optical rotatory dispersion (TRMORD) spectroscopies to the study of function and folding in heme proteins. The novel optical methods employed use quasi-null ellipsometry and polarimetry to study rapid kinetic processes (nanosecond to second time scales) in biomolecules that contain magneto-optically active chromophores such as heme and the aromatic amino acids. The overall program of the functional ligand-rebinding studies is to investigate protein relaxation after ligand photolysis with the goal of understanding how protein structure modulates the reactivity of the heme prosthetic group and elicits the variety of functions that heme proteins perform in oxidative metabolism. The proposed TRMCD studies of kinetic intermediates in the oxygen transport protein hemoglobin have the goal of better understanding the dynamics of the allosteric R yields T transition in this cooperative system. The TRMCD spectra of the near-UV tryptophan bands and the heme-based Soret and visible bands will be examined for MCD transients on the nanosecond and microsecond timescales that are characteristic of globin tertiary and quaternary structural changes after photolysis of the R-state carboxy adduct. Similarly, TRMCD/MORD studies of ligand photolysis in myoglobin will be directed toward resolving outstanding questions about the heme pocket dynamics underlying the function of this oxygen storage protein. Molecular oxygen is ultimately consumed in cells by redox reactions catalyzed at a copper-heme iron site in the enzyme cytochrome c oxidase. The TRMCD studies of ligand dynamics at the bimetallic site proposed here are ultimately addressed at a major puzzle in biophysics: How does cytochrome oxidase couple the energy released in this redox chemistry to the pumping of protons against a gradient? Finally, in the other major direction of investigation proposed, TRMCD/MORD techniques will be used to monitor ultrafast (submillisecond) events in the folding reactions of heme proteins. In particular, this work will look for direct spectrokinetic evidence for the type of biased diffusional dynamics thought to characterize the earliest events in the folding of protein chains in the new, energy landscape point of view.